蜡梅小孢子发生和花粉形成的研究

通过对蜡梅小孢子发生和花粉形成的研究,结果表明：蜡梅幼小花药中的多列孢原细胞经造孢细胞发良为小孢子母细胞。减数分裂为同时型。四分体呈四面体型排列,同时观察了小孢子在发育过程中液泡的动态变化。成熟花粉为２－细胞型。花药壁的发育为双子叶型。花药壁由５－６层细胞组成,腺质绒毡层。花粉具有异型性现象。     

羽叶薰衣草雄性不育的细胞学研究

用光镜和电镜观察羽叶薰衣草(Lavandula pinnata L.)雄性不育小孢子发育过程的细胞形态学特征.结果表明:羽叶薰衣草花药4枚,每枚花药通常具4个小孢子囊.花药壁发育为双子叶型,从外向内分为表皮、药室内壁、中层和绒毡层4层细胞.减数分裂形成的四分体为四面体及十字交叉型.小孢子的发育过程可分为造孢细胞期、减数分裂时期、小孢子发育早期、小孢子发育晚期.未观察到二胞花粉期和成熟花粉期.羽叶薰衣草花粉败育主要发生在单核花粉时期,细胞内物质解体并逐渐消失变成空壳花粉或花粉皱缩变形成为各种畸形的败育花粉.在此之前小孢子的发育正常.羽叶薰衣草小孢子不育机制体现在绒毡层过早解体、四分体时期以后各细胞中线粒体结构不正常、胼胝质壁与小孢子母细胞脱离、花药壁细胞中淀粉出现时间异常等. 壁发育为双子叶型,从外向内分为表皮、药室内壁、中层和绒毡层4层细胞.减数分裂形成的四分体为四面体及十字交叉型.小孢子的发育过程可分为造孢细胞期、减数分裂时期、小孢子发育早期、小孢子发育晚期.未观察到二胞花粉期和成熟花粉期.羽叶薰衣草花粉败育主要发生在单核花粉时期,细胞内物质解体并逐渐消失变成空壳花粉或花粉皱缩变形成为各种畸形的败育花粉.在此 前小孢子的发育正常.羽叶薰衣草小孢子不育机制体现在绒毡层过早解体、四分体时期以后各细胞中线粒体结构不正常、胼胝质壁与小孢子母细胞脱离、花药壁细胞中淀粉出现时间异常等. 壁发育为双子叶型,从外向内分为表皮、药室内壁、中层和绒毡层4层细胞.减数分裂形成的四分体为四     

侧柏小孢子的发生和雄配子体的形成

侧柏[Platycladusorientalis（L．）Franco]初生造孢细胞在8月下旬（1992年）形成,11月上旬形成小孢子母细胞,1993年2月中旬形成四分体,2月下旬小孢子从四分体内释放出来,3月中旬形成成熟花粉粒并开始传粉,4月上旬花粉粒在珠心上萌发,5月上旬生殖细胞分裂,6月上旬精原细胞分裂。小孢子母细胞在休眠以前开始减数分裂,解除休眠以后形成成熟的花粒粒。减数分裂从11月上旬开始至次年2月17日结束。小孢子母细胞减数分裂存在扩散双线期。小孢子母细胞以双线期渡过休眠。精原细胞接近颈卵器时开始分裂,形成两个大小相同的精细胞。精细胞独立存在的时间很短。精细胞的细胞质分为三个区域。小孢子母细胞在发育过程中,发现有部分小孢子母细胞退化,在小孢子囊内形成一大的空腔的现象。     

高羊茅花粉母细胞减数分裂及其雄性不育的细胞生理学研究

为探讨高羊茅雄性不育株A22013189的小孢子发育过程及其败育的生理学机理,以可育株189为对照,对其结实能力、花粉活力、花粉母细胞减数分裂进程中的染色体行为及生理生化特征进行研究。结果表明:(1)不育株A22013189花粉数量少,花粉粒空瘪皱褶,自交不结实,在杂交中作父本获得杂交后代的可能性极小。(2)从减数分裂前期Ⅰ至四分体时期,不育株A22013189花粉母细胞存在大量落后染色体、染色体桥和断片、微核、单价染色体、不均等分离、染色体分裂不同步、游离染色体、三分体、60°纺锤体、染色体缺失等异常现象,初步分析这些小孢子异常分裂是导致高羊茅花粉败育的细胞学原因之一。(3)不育株A22013189的可溶性蛋白质、可溶性糖含量在整个发育时期都显著低于同期可育株189;苗期至造孢细胞期,A22013189游离脯氨酸含量与可育株189无显著差异,但减数分裂期至花粉成熟期,A22013189游离脯氨酸含量却显著低于同期可育株189;苗期至造孢细胞期,A22013189丙二醛含量显著低于同期可育株189,但小孢子进入减数分裂期后,A22013189丙二醛的增加速度和积累量明显高于同期可育株189。研究发现,高羊茅雄性不育株花粉母细胞减数分裂染色体行为异常,生长发育过程存在物质能量代谢降低,有害物质积累现象。研究结果对于高羊茅败育机理研究及杂交育种亲本选择有重要的理论指导意义。     

越南篦齿苏铁小孢子发生及其系统学意义

运用常规石蜡切片方法,结合显微荧光技术对越南篦齿苏铁Cycas elongata 小孢子发生和花粉个体发育进行了研究。结果表明：其小孢子叶球5月中下旬开始萌动,小孢子囊着生在小孢子叶远轴面,且3-5小孢子囊以辐射状排列方式聚生成聚合囊。小孢子囊壁由6-7层细胞组成,包括表皮、中层及绒毡层。绒毡层来源于成熟造孢组织的外围细胞,其退化形式为分泌型。6月中旬,小孢子母细胞进入减数分裂I,至6月下旬形成四分体。母细胞减数分裂后胞质分裂的方式与其他苏铁类植物不同,具有连续型与同时型两种类型。7月中旬,小孢子经过2次有丝分裂后,形成3细胞的成熟花粉粒。7月下旬进入散粉状态。在花粉发育过程中,母细胞内淀粉粒的积累及其壁上胼胝质的沉积均呈现规律性变化。     

南方红豆杉小孢子发生与雄配子体发育

采用石蜡切片法,对南方红豆杉小孢子发生及雄配子体发育过程进行了系统地观察。结果表明：南方红豆杉小孢子叶球于7月下旬分化,9月中旬形成造孢细胞,11月初形成小孢子母细胞;同一小孢子叶球中的小孢子母细胞表现出发育不同步现象;11中旬,进入减数分裂时期,形成游离小孢子后休眠越冬,于翌年1月下旬逐渐成熟,成熟花粉粒为单核;2月中下旬开始散粉,散粉时间持续15 d 左右。花粉落入胚珠后,经过3次分裂形成管细胞、柄细胞和2个精子;管细胞和柄细胞最终退化解体,未见花粉败育现象。认为南方红豆杉小孢子发生与雄配子体发育正常,不是致其濒危的主要原因。     

观光木的小孢子发生及雄配子体发育的研究

采用石蜡切片法对观光木(Tsoongiodendron odorum Chun)的小孢子发生和雄配子体发育进行了解剖学研究.观光木的花药由花药原基发育而来,具4个小孢子囊,花药壁由表皮、药室内壁、2～3层中层和1～2层绒毡层组成.中层在小孢子四分体时期开始解体,最终消失;绒毡层为腺质绒毡层,细胞具1～2核,在花药发育过程中不断分泌各种物质,提供小孢子发育,直到花粉成熟绒毡层才自溶消失.初生造孢细胞分裂形成次生造孢细胞,次生造孢细胞再转化为小孢子母细胞,小孢子母细胞减数分裂的胞质分裂为修饰性同时型,四分体排列方式为交叉型、对称型或"T"型(极少),成熟花粉粒二细胞型,开花时散出.观光木的成熟花粉粒存在严重的败育现象.     

革苞菊胚胎学研究 Ⅰ.大、小孢子发生和雌、雄配子体发育

革苞菊为雌雄异株。在雄花中 ,花药 4室 ,药壁发育为双子叶型 ,由表皮、药室内壁 ,一层中层和绒毡层组成。绒毡层于小孢子四分体时期开始变形 ,其细胞原生质体向药室中移动 ,为变形绒毡层。小孢子孢原为多细胞 ,小孢子母细胞减数分裂产生四面体型的小孢子四分体。四分体胞质分裂为同时型。成熟花粉 3-细胞型。单核期的小孢子出现壁发育不良和巨大及空花粉现象。在雌花中 ,胚珠是倒生的 ,单珠被 ,薄珠心 ,珠被于孢原期已发育完整。大孢子孢原单细胞。由孢原细胞直接发育形成大孢子母细胞。 4个大孢子直线型 ,蓼型胚囊。于成熟胚囊期观察到发育异常的胚囊。通过对胚囊发育过程中营养物质消长规律的研究 ,讨论了环境与发育的相关性问题。     

山麦冬大小孢子发生及雌雄配子体发育研究

采用石蜡切片技术对百合科植物山麦冬大小孢子发生及雌雄配子体发育进行了观察研究。结果表明:(1)山麦冬花药具有4个花粉囊,花药壁的发育方式为基本型,花药壁完全分化时由表皮、药室内壁、中层及绒毡层组成。(2)绒毡层发育类型为分泌型,到四分体孢子彼此分离形成单细胞花粉阶段,绒毡层细胞开始解体退化,花粉成熟时绒毡层细胞完全消失;花粉母细胞减数分裂为连续型,四分体为左右对称形排列,成熟花粉为3-细胞花粉,单萌发沟。(3)子房3室,每室2枚胚珠,胚珠倒生型,双珠被,薄珠心,雌性孢原细胞不经过平周分裂而直接发育而成大孢子母细胞。(4)减数分裂后四分体大孢子呈线型或T型排列,合点端大孢子分化为功能大孢子,胚囊发育为蓼型;花粉母细胞减数分裂过程中,二分体、四分体细胞外方被胼胝质壁所包被,小孢子形成后胼胝质壁逐渐消失。该研究结果丰富了百合科植物生殖生物学研究的内容,也为探讨百合科植物的系统学研究提供了参考。     

水稻小孢子发育过程中微管骨架的变化

对水稻（ＯｒｙｚａｓａｔｉｖａＬ．）小孢子发育过程中微管变化的研究表明,微管在小孢子不同发育阶段呈现多样性。在花粉母细胞内,微管形成许多粗束和分支,围绕着核分布形成一个网络。花粉母细胞经第一次减数分裂形成二分体。在每一个二分体细胞内,有许多微管束,从核周辐射至细胞质各部位;在细胞质存在一个疏松的微管束网络。二分体经第二次减数分裂形成四分体,在每一个四分体细胞内,微管束呈辐射状,从核膜辐射入细胞质内。四分体形成后不久,四分体的四个细胞便分开,每一个细胞变成一个独立的小孢子。在早期的小孢子细胞内,微管束呈疏松网状分布。其中有些微管束朝向胞质一个小突起聚集。当小孢子进入中期发育阶段,在胞质的小突起部位微管束密度增大。小突起最终形成为萌发孔。当小孢子发育至成熟期,细胞内的微管束变得纤细,而网络则变得紧密。之后的发育阶段（即花粉发育不同阶段）因荧光标记难以进入细胞,无法获得清晰的图像。     

Ultrastructure of microsporogenesis and microgametogenesis in Campsis radicans (L.) Seem. (Bignoniaceae)

In the present study, microsporogenesis, microgametogenesis and pollen wall ontogeny in Campsis radicans (L.) Seem. were studied from sporogenous cell stage to mature pollen using transmission electron microscopy. To observe the ultrastructural changes that occur in sporogenous cells, microspores and pollen through progressive developmental stages, anthers at different stages of development were fixed and embedded in Araldite. Microspore and pollen development in C. radicans follows the basic scheme in angiosperms. Microsporocytes secrete callose wall before meiotic division. Meiocytes undergo meiosis and simultaneous cytokinesis which result in the formation of tetrads mostly with a tetrahedral arrangement. After the development of free and vacuolated microspores, respectively, first mitotic division occurs and two-celled pollen grain is produced. Pollen grains are shed from the anther at two-celled stage. Pollen wall formation in C. radicans starts at tetrad stage by the formation of exine template called primexine. By the accumulation of electron dense material, produced by microspore, in the special places of the primexine, first of all protectum then columellae of exine elements are formed on the reticulate-patterned plasma membrane. After free microspore stage, exine development is completed by the addition of sporopollenin from tapetum. Formation of intine layer of pollen wall starts at the late vacuolated stage of pollen development and continue through the bicellular pollen stage.     

凤仙花花药发育中多糖和脂滴组织化学研究

以不同发育时期的凤仙花花药为实验材料,采用组织化学方法,对花药发育中的结构变化及多糖和脂滴物质分布进行观察。结果表明:(1)凤仙花的花药壁由6层细胞组成,包括1层表皮细胞,2层药室内壁细胞,2层中层细胞和1层绒毡层细胞。其中绒毡层细胞的形态不明显,很难与造孢细胞区分,且在小孢子母细胞时期退化。(2)在小孢子母细胞中出现了一些淀粉粒,但减数分裂后,早期小孢子中的淀粉粒消失,又出现了一些小的脂滴;随着花粉的发育,小孢子形成大液泡,晚期小孢子中的脂滴也消失;小孢子分裂形成二胞花粉后,营养细胞中的大液泡降解、消失,二胞花粉中又开始积累淀粉;接近开花时,成熟花粉中充满细胞质,其中包含了较多的淀粉粒和脂滴。(3)在凤仙花的花药发育中,绒毡层细胞很早退化,为小孢子母细胞和四分体小孢子提供了营养物质;其后的中层细胞退化则为后期花粉发育提供了营养物质。     

ULTRASTRUCTURE AND DEVELOPMENT OF THE NEXINE AND INTINE IN THE POLLEN WALL OF SILENE ALBA (CARYOPHYLLACEAE)

Nexine and intine development in Silene alba (Caryophyllaceae) was investigated by electron microscopy and enzyme cytochemistry. Nexine-2 forms by deposition of sporopollenin along unit membrane lamellae closely associated with the microspore plasma membrane in the late tetrad stage. After the callose wall dissolves, electron density increases along the tangentially oriented fibers of the proximal primexine, forming nexine-1. When the exine is essentially complete, the intine begins to develop. In the nearly mature microspore, acid phosphatase activity appears in the peripheral cytoplasm just prior to its extrusion into the intine of the mature pollen grain.     

A glycine-rich protein that facilitates exine formation during tomato pollen development

Formation of the unique and highly diverse outer cell wall, or exine, of pollen is essential for normal pollen function and survival. However, little is known about the many contributing proteins and processes involved in the formation of this wall. The tomato gene LeGRP92 encodes for a glycine-rich protein produced specifically in the tapetum. LeGRP92 is found as four major forms that accumulate differentially in protein extracts from stamens at different developmental stages. The three largest molecular weight forms accumulated during early microspore development, while the smallest molecular weight form of LeGRP92 was present in protein extracts from stamens from early microsporogenesis through anther dehiscence, and was the only form present in dehisced pollen. Light microscopy immunolocalization experiments detected LeGRP92 at only two stages, late tetrad and early free microspore. However, we observed accumulation of the LeGRP92 at the early tetrad stage of development by removing the callose wall from tetrads, which allowed LeGRP92 detection. Transmission electron microscopy confirmed the LeGRP92 accumulation from microspore mother cells, tetrads through anther dehiscence. It was observed in the callose surrounding the microspore mother cells and tetrads, the exine of microspores and mature pollen, and orbicules. Plants expressing antisense RNA had reduced levels of LeGRP92 mRNA and protein, which correlated to pollen with altered exine formation and reduced pollen viability and germination. These data suggest that the LeGRP92 has a role in facilitating sporopollenin deposition and uniform exine formation and pollen viability.     

Distribution of insoluble polysaccharides, neutral lipids, and proteins in the developing anthers of Campsis radicans (L.) Seem. (Bignoniaceae)

In this study, distribution of polysaccharides, lipids, and proteins in the developing anthers of Campsis radicans (L.) Seem. was examined from sporogenous cell stage to mature pollen, using cytochemical methods. To detect the distribution and dynamic changes of insoluble polysaccharides, lipid bodies, and proteins in the anthers through progressive developmental stages, semi-thin sections of anthers at different developmental stages were stained with periodic-acid-Schiff (PAS) reagent, Sudan black B, and Coomassie brilliant blue, respectively, and examined under light microscope. Ultrastructural observations with TEM were also carried out to determine the storage form of starch in the connective tissue, and storage form of lipids in the tapetal cells. In sporogenous cell stage, anther wall contains numerous insoluble polysaccharides. However, from the sporogenous cell stage to the vacuolated microspore stage, the amount of insoluble polysaccharides in the anther wall decreases gradually. At bicellular pollen stage, tapetum degenerates completely and polysaccharides are not seen in the anther wall. Lipid bodies are observed in the cytoplasm of both middle layer and tapetal cells at tetrad stage, whereas they disappear in the vacuolated microspore stage. Compared with polysaccharides, proteins are limited in the anther wall at early stages of development. During pollen development, polysaccharides, proteins, and lipid bodies are scarce in the cytoplasm of sporogenous cells, but their amount increases at premeiotic stage. From tetrad stage to bicellular pollen stage, microspore cytoplasm contains variable amount of insoluble polysaccharide grains, lipid and protein bodies. At bicellular pollen stage, plentiful amount of starch granules are stored in the cytoplasm of the pollen grains. Proteins and lipid bodies are also present in the cytoplasm.     

车桑子小孢子发生与雄配子体发育

为了解干热河谷区车桑子(Dodonaea viscosa)胚胎学特征及其结籽率低的原因,采用常规石蜡切片法和电镜扫描技术对车桑子小孢子发生、雄配子体发育和花粉的形态特征进行了观察。结果表明,车桑子花药具有4个花粉囊。完整的花药壁从外到内依次为表皮、药室内壁、2～3层中层细胞和绒毡层;绒毡层类型是腺质绒毡层。花药成熟期,中层、绒毡层均退化消失。小孢子母细胞进行同时型胞质分裂;四分体为四面体型结构。成熟的花粉为二细胞型。花粉近球形,外壁密布颗粒状纹饰,具有3条不构成合沟的萌发沟。雄性生殖发育过程出现的异常现象可能是干热河谷地区车桑子结籽率低的原因之一。     

Amorphophallus: New insights into pollen morphology and the chemical nature of the pollen wall

In pollen characters, Amorphophallus is one of the most diverse genera in the Araceae. The present work is a critical survey of contradicting reports on the impact of acetolysis treatment on Amorphophallus pollen, on the chemical nature of the outer pollen wall layer and of electron-dense (dark) granules found within it. Furthermore, we wanted to clarify the pollen polarity and to test conclusions based on different preparation techniques. Pollen morphology of 25 species is investigated by light microscopy, scanning electron microscopy and transmission electron microscopy. Our results show that Amorphophallus pollen is not resistant to acetolysis treatment. The use of different transmission electron microscopy staining methods proved the polysaccharide nature of the outer pollen wall layer and of the granules within it. Moreover, an additional thin surface layer was found in all investigated species. Microspores in early and late tetrad stages show that the less convex side of the microspore is the proximal face and the more convex side the distal face. The extrusion of pollen in strands is illustrated for the first time by light microscopy and scanning electron microscopy. Furthermore, observations of pollen in water showed that in some of the investigated species the pollen wall is shed immediately before pollen tube formation.     

DEVELOPMENT OF OMNIAPERTURATE POLLEN IN TRILLIUM KAMTSCHATICUM (LILIACEAE)

Ultrastructural changes during omniaperturate pollen development in Trillium kamtschaticum Pall, was examined using transmission electron microscopy. The pollen mother cells are not enveloped within a thick callosic wall. The microspores resulting from successive meiosis are divided by scanty deposition of callosic wall in the tetrad. A primexine/exine template is not recognizable within the tetrad during formation of exinous components. Preexinous globules, originating from vesicles in the callosic wall, accumulate electron-dense materials and develop into exinous globules. The preexinous globules have ca 10 nm wide contacts with tilted and invaginated plasma membrane of the microspore within the callosic wall. After dissolution of the callosic wall, the microspores separate and mitosis subsequently leads to the formation of a generative cell and vegetative cell encased in a loose aggregation of developing exinous globules. When the generative cell is at the pollen grain surface, the channeled zone is initiated at the opposite side of the microspore on the surface of the vegetative cell. Just before pollen maturity, a new layer develops under the channeled zone. Thus, development of the omniaperturate pollen grains of T. kamtschaticum involves some processes that are distinct from those of Canna and Heliconia and some that are similar.     

Anther ontogeny in <Emphasis Type="Italic">Brachypodium distachyon</Emphasis>

Brachypodium distachyon has emerged as a model plant for the improvement of grain crops such as wheat, barley and oats and for understanding basic biological processes to facilitate the development of grasses as superior energy crops. Brachypodium is also the first species of the grass subfamily Pooideae with a sequenced genome. For obtaining a better understanding of the mechanisms controlling male gametophyte development in B. distachyon, here we report the cellular changes during the stages of anther development, with special reference to the development of the anther wall. Brachypodium anthers are tetrasporangiate and follow the typical monocotyledonous-type anther wall formation pattern. Anther differentiation starts with the appearance of archesporial cells, which divide to generate primary parietal and primary sporogenous cells. The primary parietal cells form two secondary parietal layers. Later, the outer secondary parietal layer directly develops into the endothecium and the inner secondary parietal layer forms an outer middle layer and inner tapetum by periclinal division. The anther wall comprises an epidermis, endothecium, middle layer and the secretory-type tapetum. Major documented events of anther development include the degradation of a secretory-type tapetum and middle layer during the course of development and the rapid formation of U-shaped endothecial thickenings in the mature pollen grain stage. The tapetum undergoes degeneration at the tetrad stage and disintegrates completely at the bicellular stage of pollen development. The distribution of insoluble polysaccharides in the anther layers and connective tissue through progressive developmental stages suggests their role in the development of male gametophytes. Until sporogenous cell stage, the amount of insoluble polysaccharides in the anther wall was negligible. However, abundant levels of insoluble polysaccharides were observed during microspore mother cell and tetrad stages and gradually declined during the free microspore and vacuolated microspore stages to undetectable level at the mature stage. Thus, the cellular features in the development of anthers in B. distachyon share similarities with anther and pollen development of other members of Poaceae.